A Narrative Review of Parameters Influencing Preeclampsia in the COVID-19 Era.

The COVID-19 outbreak has emerged as one of the most profound medical events of the 21st century, leaving an indelible impact on a global scale. The widespread prevalence causing significant illness and death needs collaborative and inventive efforts to deal with this challenge. One of the particular subset of the general population that had endured a significant impact was the pregnant population. A key complication of pregnancy seen in patients with a COVID-19 infection was the increased risk of developing preeclampsia. The angiotensin-converting enzyme 2 (ACE2) receptor is an important part of the renin-angiotensin system, which has been implicated in the control of blood flow and also is a key receptor in the pathogenesis of the multitude of symptoms of COVID-19. This study aimed to evaluate the psychiatric, hematological, neurological, and social factors influenced by the COVID-19 virus and its subsequent effect on the development of preeclampsia. Increased rates of anxiety and depression were seen globally during the COVID-19 pandemic and due to the following physiological response of anxiety and depression, elevated blood pressure levels and development of preeclampsia were noted. Neurological factors such as the development of posterior reversible encephalopathy syndrome and its relationship between COVID-19 and preeclampsia were also strongly observed. The observation suggested biomarkers such as serum neurofilament light may be used as a screening tool to stratify the severity of preeclampsia. Hematological parameters observed were most notable for the presence of thrombocytopenia, which itself is a marker of the severity of preeclampsia. The numerous effects of COVID-19 on preeclampsia have proven to have a tremendous impact on the healthcare burden. Careful analysis and prevention strategies, if implemented, will contribute to reducing the morbidity and mortality of patients with preeclampsia and COVID-19 infections.

Factors in the Development of Somatoform Disorders Among Children: A Case-Control Study.

Introduction Somatoform disorders (SFDs) are a spectrum of diseases mainly manifesting with physical symptoms of no recognizable etiology. These disorders are believed to be primarily influenced and exacerbated by psychological factors. Given the connection between parental sociodemographics and psychological factors and SFDs, there is a pressing need to investigate this area further, particularly concerning parents and their children affected by somatoform disorders. Aims and objectives The aims and objectives of this study are as follows: i) study the determinants of SFDs, namely, parent handling of child, parent-child relationship, parenting with respect to attending to the needs of children, and intelligence quotient (IQ) of parents, and ii) compare host factors to the factors matched in control subjects. Materials and methods We adopted purposive sampling in our case-control study. The study sample was obtained from the psychiatry department of the Ganesh Shankar Vidyarthi Memorial (GSVM) Medical College, Kanpur, India, from September 2020 to December 2022, once a week, every Monday. Children manifesting SFD manifestations that are among the chief complaints were included in our study. Results A total of 115 cases were included in our study based on inclusion criteria. The study compared sociodemographic characteristics, intelligence quotient (IQ), parental characteristics, parental handling, parent-child relationships, and parenting practices between a patient group and a control group. The results showed no significant differences in sex, religion, occupation, domicile, and socioeconomic status in both control and patient groups. However, significant differences were found in parental characteristics, such as lower mean age, education, and IQ, in the patient group. This difference between the patient and control groups with respect to the parental handling questionnaire was statistically significant for the domain of "praise," "talk," "feel better," "comes to you," "unduly strict" items, "frequently reprimanded," "tried to control everything," creative activities, protectiveness, education, neglecting, objective punishment, housing, medical care, demanding, symbolic reward, loving, objective reward, food, parent-to-child communication, clothing, support, routine, recreation, social activities, rules, managing problem behavior, guidance for career, and security. Conclusion Parents were deficient in terms of years of education, occupational status, IQ, parental handling, parent-child relationship, and parenting with respect to the children's needs. These findings offer insights into the sociodemographic and psychological factors contributing to the patient group's condition.

Use of Drugs Considering Route of Administration on Drug Prescription Pattern for Bronchial Asthma in a Tertiary Level Hospital.

Bronchial asthma is one of the much known long-term respiratory conditions. Incidence is increasing, in developing countries like Bangladesh. Cross-sectional type of observational study was carried out over one year (July 2017 to June 2018) in the department of Pharmacology with collaboration of the department of Respiratory Medicine and Medicine, Mymensingh Medical College and Hospital, Mymensingh, Bangladesh. A total of 160 patients were selected non-randomly for the study. Inhalation route (52.35%) was the most preferred one over oral route (47.65%). In total 245 drugs, 131 FDC drugs (Salmeterol + Fluticasone) were prescribed with inhalation therapy which is 53.46%, another 9 FDC drugs that is (Ipratropium bromide + Salbutamol) were prescribed with inhalation therapy which is 3.67%, 101 drugs (Salbutamol) were prescribed with inhalation therapy that is 41.23%, 4 drugs (Beclomethasone) were prescribed with inhalation therapy that is 1.64%. Majority of patient were taking inhalation form of anti-asthmatic drugs.

Interaction of Eating Status and Dietary Variety on Incident Functional Disability among Older Japanese Adults.

OBJECTIVES: To examine whether eating status and dietary variety were associated with functional disability during a 5-year follow-up analysis of older adults living in a Japanese metropolitan area. DESIGN: A 5-year follow-up study. SETTING: Ota City, Tokyo, Japan. PARTICIPANTS: A total of 10,308 community-dwelling non-disabled adults aged 65-84 years. MEASUREMENTS: Eating status was assessed using a self-reported questionnaire. Dietary variety was assessed using the dietary variety score (DVS). Based on the responses, participants were classified according to eating alone or together and DVS categories (low: 0-3; high: 4-10). Functional disability incidence was prospectively identified using the long-term care insurance system's nationally unified database. Multilevel survival analyses calculated the adjusted hazard ratio (HR) and 95% confidence interval (CI) for incident functional disability. RESULTS: During a 5-year follow-up, 1,991 (19.3%) individuals had functional disabilities. Eating status or DVS were not independently associated with incident functional disability. However, interaction terms between eating status and DVS were associated with functional disability; HR (95% CI) for eating together and low DVS was 1.00 (0.90-1.11), eating alone and high DVS was 0.95 (0.77-1.17), and eating alone and low DVS was 1.20 (1.02-1.42), compared to those with eating together and high DVS. CONCLUSION: Older adults should avoid eating alone or increase dietary variety to prevent functional disability. This can be ensured by providing an environment of eating together or food provision services for eating a variety of foods in the community.

Score-Based and Nutrient-Derived Dietary Patterns Are Associated with Depressive Symptoms in Community-Dwelling Older Japanese: A Cross-Sectional Study.

OBJECTIVES: This study evaluated associations of score-based and nutrient-derived dietary patterns with depressive symptoms in community-dwelling older Japanese. DESIGN: Cross-sectional study. SETTING: Community-based. PARTICIPANTS: 982 community-dwelling adults aged 65 years or older. MEASUREMENTS: Score-based pattern was assessed by using dietary variety score (DVS), which covers 10 food group items in Japanese meals. Nutrient-derived dietary patterns were identified by using reduced rank regression (RRR), with folate, vitamin C, magnesium, calcium, iron, and zinc intakes as response variables. Depressive symptoms were assessed with the Geriatric Depression Scale. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated for these dietary patterns in multivariate logistic regression analyses with potential confounders. The lowest consumption category was used as the reference group. RESULTS: The prevalence of depressive symptoms was 13.5%. Higher DVS was associated with fewer depressive symptoms (OR=0.52, 95% CI=0.27-1.03 for the highest vs the lowest DVS; P for trend=0.031). The first RRR dietary pattern score was characterized by high intakes of fish, soybean products, potatoes, most vegetables, mushrooms, seaweeds, fruits, and green tea and a low intake of rice and was inversely associated with the prevalence of depressive symptoms (OR=0.53, 95% CI=0.30-0.92; P for trend=0.030). CONCLUSION: Greater dietary variety and a dietary pattern characterized by high intakes of fish, soybean products, potatoes, most vegetables, mushrooms, seaweeds, fruit, and green tea and a low intake of rice were consistently associated with lower prevalence of depressive symptoms in community-dwelling older Japanese. Therefore, both patterns identified the components of dietary habits essential to depression prevention.

Antibacterial Effect of Aloe vera (Aloe barbadensis) leaf gel against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae.

This experimental study was carried out to determine the antibacterial effect of Ethanolic extract of Aloe vera leaf gel (EAE) against standard strains of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae in the Department of Pharmacology & Therapeutics in collaboration with the Department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh from July 2017 to June 2018. Ethanol extract was used in five different concentrations (100, 200, 300, 400 and 500µg/ml). Dose dependent inhibitory effect was seen against the test organisms using disc diffusion method. For S. aureus, Zones of inhibition (ZOI) were 7, 12, 13, 16 and 20 mm at 100, 200, 300, 400 and 500 µg/ml respectively; for P. aeruginosa, ZOI were 0, 7, 12, 14 and 17 mm at 100, 200, 300, 400 and 500 µg/ml respectively; for E. coli, ZOI were 0, 8, 12, 15 and 18 mm at 100, 200, 300, 400 and 500 µg/ml respectively; for K. pneumoniae, ZOI were 7, 10, 11, 13 and 17 mm at 100, 200, 300, 400 and 500 µg/ml respectively. The minimum inhibitory concentration (MIC) was assessed by broth dilution technique. The MICs of EAE for S. aureus, P. aeruginosa, E. coli and K. pneumoniae were 500, 650, 650 and 600 µg/ml respectively. From the study it is clearly observed that ethanolic extract of Aloe vera leaf gel possesses antibacterial effect against the test pathogens. Further studies are required to detect and isolate the biologically active ingredients present in the Aloe vera leaves which are responsible for this antibacterial effect. Hopefully, that would lead to the discovery of new and more potent antimicrobial agents originated from Aloe vera.

Identification of the first homozygous 1-bp deletion in GDF9 gene leading to primary ovarian insufficiency by using targeted massively parallel sequencing.

Targeted massively parallel sequencing (TMPS) has been used in genetic diagnosis for Mendelian disorders. In the past few years, the TMPS has identified new and already described genes associated with primary ovarian insufficiency (POI) phenotype. Here, we performed a targeted gene sequencing to find a genetic diagnosis in idiopathic cases of Brazilian POI cohort. A custom SureSelectXT DNA target enrichment panel was designed and the sequencing was performed on Illumina NextSeq sequencer. We identified 1 homozygous 1-bp deletion variant (c.783delC) in the GDF9 gene in 1 patient with POI. The variant was confirmed and segregated using Sanger sequencing. The c.783delC GDF9 variant changed an amino acid creating a premature termination codon (p.Ser262Hisfs*2). This variant was not present in all public databases (ExAC/gnomAD, NHLBI/EVS and 1000Genomes). Moreover, it was absent in 400 alleles from fertile Brazilian women screened by Sanger sequencing. The patient's mother and her unaffected sister carried the c.783delC variant in a heterozygous state, as expected for an autosomal recessive inheritance. Here, the TMPS identified the first homozygous 1-bp deletion variant in GDF9. This finding reveals a novel inheritance pattern of pathogenic variant in GDF9 associated with POI, thus improving the genetic diagnosis of this disorder.

Albumin, Hemoglobin, and the Trajectory of Cognitive Function in Community-Dwelling Older Japanese: A 13-Year Longitudinal Study.

BACKGROUND: Cognitive function can substantially decline over a long period, and understanding the trajectory of cognitive function is important. However, little is known about the linkage between nutritional biomarkers and long-term cognitive change. OBJECTIVES: We analyzed 13-year longitudinal data for older Japanese to examine the associations of serum albumin and hemoglobin levels with the trajectory of cognitive function. DESIGN: Longitudinal study. SETTING: Community-based. PARTICIPANTS: A total of 1,744 community-dwelling adults aged 65 years or older who participated in annual health examinations in Kusatsu town, Gunma Prefecture, Japan, from 2002-2014. MEASUREMENTS: Cognitive function was assessed annually by the Mini-Mental State Examination (MMSE). Albumin and hemoglobin levels at baseline (the year when a respondent first participated in the health examination) were divided into quartiles. Hierarchical linear modeling was used to analyze intrapersonal and interpersonal differences in cognitive function. RESULTS: Participants' MMSE scores decreased at an accelerated rate over the 13-year period. Participants with the lowest baseline albumin level (below the first quartile line) showed a greater accelerated decline in MMSE scores over time, compared with those with the highest level (above the third quartile line). Moreover, MMSE scores in participants with a lower hemoglobin level and lower MMSE score at baseline tended to decline faster over time at an accelerated rate. CONCLUSIONS: These findings yield new insights about the complex and diverse roles of these nutritional biomarkers on the trajectory of cognitive function in old age.

Malignant testicular germ cell tumors in postpubertal individuals with androgen insensitivity: prevalence, pathology and relevance of single nucleotide polymorphism-based susceptibility profiling.

STUDY QUESTION: What is the prevalence of malignant testicular germ cell tumors (TGCT) and its precursors, (pre-) germ cell neoplasia in situ (GCNIS), in late teenagers and adults who have androgen insensitivity syndrome (AIS) and the impact of an individual's genetic susceptibility to development of TGCT? SUMMARY ANSWER: No GCNIS or TGCT was diagnosed, but pre-GCNIS was identified in 14 and 10% of complete and partial AIS patients, respectively, and was associated with a higher genetic susceptibility score (GSS), with special attention for KITLG (rs995030) and ATFZIP (rs2900333). WHAT IS KNOWN ALREADY: Many adult women with AIS decline prophylactic gonadectomy, while data regarding the incidence, pathophysiology and outcomes of TGCT in postpubertal individuals with AIS are lacking. The relevance of genetic factors, such as single nucleotide polymorphisms (SNPs), in predisposing AIS individuals to TGCT is unknown. STUDY DESIGN, SIZE, DURATION: This multicenter collaborative study on prophylactically removed gonadal tissue was conducted in a pathology lab specialized in germ cell tumor biology. PARTICIPANTS/MATERIALS, SETTING, METHODS: Material from 52 postpubertal individuals with molecularly confirmed AIS (97 gonadal samples) was included; the median age at surgery was 17.5 (14-54) years. Immunohistochemical studies and high-throughput profiling of 14 TGCT-associated SNPs were performed. The main outcome measures were the prevalence of pre-GCNIS, GCNIS and TGCT, and its correlation with a GSS, developed based on the results of recent genome-wide association studies. MAIN RESULTS AND ROLE OF CHANCE: The earliest recognizable change preceding GCNIS, referred to as pre-GCNIS, was present in 14% of individuals with complete and 10% of those with partial AIS at a median age of 16 years. No GCNIS or invasive TGCT were found. The median GSS was significantly greater for those with, compared to those without, pre-GCNIS (P = 0.01), with an overlap between groups. Our data suggest important roles for risk alleles G at KITLG (rs995030) and C at ATFZIP (rs2900333), among the 14 studied TGCT-associated SNPs. LARGE SCALE DATA: N/A. LIMITATIONS REASONS FOR CAUTION: A limited number of cases were included. WIDER IMPLICATIONS OF THE FINDINGS: Our data suggest that the prevalence of pre-GCNIS in individuals with AIS beyond puberty is around 15%. Genetic susceptibility likely contributes to pre-GCNIS development in AIS but factors related to malignant progression remain unclear. Although data in older patients remain scarce, malignant progression appears to be a rare event, although the natural history of the premalignant lesion remains unknown. Therefore, the practice of routine prophylactic gonadectomy in adults with AIS appears questionable and the patient's preference, after having been fully informed, should be decisive in this matter. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by research grants from the Research Foundation Flanders (FWO) (to M.C.), the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq G0D6713N) (to B.B.M. and M.C.) and the European Society for Pediatric Endocrinology (ESPE), granted by Novo Nordisk AB (to J.K.). There are no competing interests.

Melting temperatures of MgO under high pressure by micro-texture analysis.

Periclase (MgO) is the second most abundant mineral after bridgmanite in the Earth's lower mantle, and its melting behaviour under pressure is important to constrain rheological properties and melting behaviours of the lower mantle materials. Significant discrepancies exist between the melting temperatures of MgO determined by laser-heated diamond anvil cell (LHDAC) and those based on dynamic compressions and theoretical predictions. Here we show the melting temperatures in earlier LHDAC experiments are underestimated due to misjudgment of melting, based on micro-texture observations of the quenched samples. The high melting temperatures of MgO suggest that the subducted cold slabs should have higher viscosities than previously thought, suggesting that the inter-connecting textural feature of MgO would not play important roles for the slab stagnation in the lower mantle. The present results also predict that the ultra-deep magmas produced in the lower mantle are peridotitic, which are stabilized near the core-mantle boundary.

Structural Reconstruction of the Perivascular Space in the Adult Mouse Neurohypophysis During an Osmotic Stimulation.

Oxytocin (OXT) and arginine vasopressin (AVP) neuropeptides in the neurohypophysis (NH) control lactation and body fluid homeostasis, respectively. Hypothalamic neurosecretory neurones project their axons from the supraoptic and paraventricular nuclei to the NH to make contact with the vascular surface and release OXT and AVP. The neurohypophysial vascular structure is unique because it has a wide perivascular space between the inner and outer basement membranes. However, the significance of this unique vascular structure remains unclear; therefore, we aimed to determine the functional significance of the perivascular space and its activity-dependent changes during salt loading in adult mice. The results obtained revealed that pericytes were the main resident cells and defined the profile of the perivascular space. Moreover, pericytes sometimes extended their cellular processes or 'perivascular protrusions' into neurohypophysial parenchyma between axonal terminals. The vascular permeability of low-molecular-weight (LMW) molecules was higher at perivascular protrusions than at the smooth vascular surface. Axonal terminals containing OXT and AVP were more likely to localise at perivascular protrusions than at the smooth vascular surface. Chronic salt loading with 2% NaCl significantly induced prominent changes in the shape of pericytes and also increased the number of perivascular protrusions and the surface area of the perivascular space together with elevations in the vascular permeability of LMW molecules. Collectively, these results indicate that the perivascular space of the NH acts as the main diffusion route for OXT and AVP and, in addition, changes in the shape of pericytes and perivascular reconstruction occur in response to an increased demand for neuropeptide release.

Dietary Variety and Decline in Lean Mass and Physical Performance in Community-Dwelling Older Japanese: A 4-year Follow-Up Study.

OBJECTIVES: To examine associations of dietary variety with changes in lean mass and physical performance during a 4-year period in an elderly Japanese population. DESIGN: Four-year prospective study. SETTING: The Hatoyama Cohort Study and Kusatsu Longitudinal Study, Japan. PARTICIPANTS: 935 community-dwelling Japanese aged 65 years or older. MEASUREMENTS: Dietary variety was assessed using a 10-item food frequency questionnaire. Body composition was determined by multifrequency bioelectrical impedance analysis, and physical performance (grip strength and usual gait speed) was measured in surveys at baseline and 4 years later. Longitudinal analysis included only participants who were originally in the upper three quartiles of lean body mass, appendicular lean mass, grip strength, and usual gait speed. The outcome measures were decline in lean body mass, appendicular lean mass, grip strength, and usual gait speed, defined as a decrease to the lowest baseline quartile level at the 4-year follow-up survey. Associations of dietary variety with the outcome measures were examined by logistic regression analysis adjusted for potential confounders. RESULTS: In the fully adjusted model, the odds ratios for decline in grip strength and usual gait speed were 0.43 (95% confidence interval, 0.19-0.99) and 0.43 (confidence interval, 0.19-0.99), respectively, for participants in the highest category of dietary variety score as compared with those in the lowest category. Dietary variety was not significantly associated with changes in lean body mass or appendicular lean mass. CONCLUSION: Among older adults, greater dietary variety may help maintain physical performance, such as grip strength and usual gait speed, but not lean mass.

Association of Dietary Variety with Body Composition and Physical Function in Community-dwelling Elderly Japanese.

OBJECTIVES: To examine the associations of dietary variety with body composition and physical function in community-dwelling elderly Japanese. DESIGN: Cross-sectional study. SETTING: Community-based. PARTICIPANTS: A total of 1184 community-dwelling elderly adults aged 65 and over. MEASUREMENTS: Dietary variety was assessed with a food frequency questionnaire (maximum, 10 points) that encompassed the 10 main food components of Japanese meals (meat, fish/shellfish, eggs, milk, soybean products, green/yellow vegetables, potatoes, fruit, seaweed, and fats/oils). Body composition was determined by multifrequency bioelectrical impedance analysis. Physical function was assessed by measuring grip strength and usual walking speed. Multiple linear regression analysis was used to examine the associations of dietary variety with body composition and physical function. RESULTS: After adjusting for potential confounders, higher dietary variety scores were independently associated with higher lean mass (ß (SE): 0.176 (0.049), p<0.001) and appendicular lean mass (ß (SE): 0.114 (0.027), p<0.001) but not with body fat mass. Elders with a higher dietary variety score had greater grip strength and faster usual walking speed (ß (SE): 0.204 (0.071), p=0.004, and ß (SE): 0.008 (0.003), p=0.012, respectively). CONCLUSION: Greater dietary variety was significantly associated with greater lean mass and better physical function in Japanese elders. The causal relationship warrants investigation in a prospective study.

Effects of Type 1 Insulin-Like Growth Factor Receptor Silencing in a Human Adrenocortical Cell Line.

Type 1 insulin-like growth factor receptor (IGF-1R) is overexpressed in a variety of human cancers, including adrenocortical tumors. The aim of the work was to investigate the effects of IGF-1R downregulation in a human adrenocortical cell line by small interfering RNA (siRNA). The human adrenocortical tumor cell line NCI H295R was transfected with 2 specific IGF1R siRNAs (# 1 and # 2) and compared with untreated cells and a negative control siRNA. IGF1R expression was determined by quantitative reverse-transcription PCR (qRTPCR) and Western blot. The effects of IGF-1R downregulation on cell proliferation and apoptosis were assessed. IGF-1R levels were significantly decreased in cells treated with IGF-1R siRNA # 1 or # 2. Relative expression of IGF1R mRNA decreased approximately 50% and Western blot analysis revealed a 30% of reduction in IGF-1R protein. Downregulation of this gene resulted in 40% reduction in cell growth in vitro and 45% increase in apoptosis using siRNA # 2. These findings demonstrate that decreasing IGF-1R mRNA and protein expression in NCI H295R cells can partially inhibit adrenal tumor cell growth in vitro. Targeting IGF1R is a promising therapy for pediatric malignant adrenocortical tumor and can still be an option for adult adrenocortical cancer based on personalized genomic tumor profiling.

Transplantation of Restored Kidneys From Unrelated Donors After Resection of Renal Cell Carcinoma: Results From 10 Patients.

PURPOSE: To relieve the chronic shortage of donor kidneys, we conducted a prospective kidney transplantation trial using kidneys removed from 10 unrelated patients (51 to 79 years of age) who had undergone nephrectomy for small renal cell carcinoma (1.5 to 3.9 cm) of low-to-moderate complexity based on RENAL (radius, exophytic/endophytic properties, nearness of tumor to the collecting system or sinus in millimeters, anterior/posterior location relative to polar lines) nephrometry (objective description helpful for operative indication and planning). METHODS: Donors were selected from among 15 patients who opted to undergo nephrectomy for small renal cell carcinoma. A total of 76 dialysis patients 34 to 85 years of age who agreed to undergo restored kidney transplantation were recruited as transplant candidates. RESULTS: In stage 1 (5 cases), high-risk patients were selected without human leukocyte antigen testing, and accelerated acute rejection occurred in 4 of 5 recipients. This trial was subsequently extended with human leukocyte antigen testing, and an additional 5 patients were enrolled in stage 2. Eight recipients, including 4 recipients with a history of renal transplantation, experienced rejection; 1 patient resumed dialysis 35 months after transplantation. The most recent serum creatinine levels ranged from 1.10 to 3.19 mg/dL in the 9 recipients with functioning grafts and from 0.84 to 4.68 mg/dL in the 10 donors. No tumor recurrence was noted at 32 to 58 months after surgery in either the recipients or the donors. CONCLUSIONS: Restored kidney transplantation using kidneys with a small renal tumor seems suitable for carefully selected high-risk recipients and, in particular, elderly kidneys can also function well. Avoiding cancer transmission, fair recipient selection, close follow-up, and a well-organized tracking system warrant further study.

Induction of cells with cancer stem cell properties from nontumorigenic human mammary epithelial cells by defined reprogramming factors.

Cancer stem cells (CSCs), a small and elusive population of undifferentiated cancer cells within tumors that drive tumor growth and recurrence, are believed to resemble normal stem cells. Although surrogate markers have been identified and compelling CSC theoretical models abound, actual proof for the existence of CSCs can only be had retrospectively. Hence, great store has come to be placed in isolating CSCs from cancers for in-depth analysis. On the other hand, although induced pluripotent stem cells (iPSCs) hold great promise for regenerative medicine, concern exists over the inadvertent co-transplantation of partially or undifferentiated stem cells with tumorigenic capacity. Here we demonstrate that the introduction of defined reprogramming factors (OCT4, SOX2, Klf4 and c-Myc) into MCF-10A nontumorigenic mammary epithelial cells, followed by partial differentiation, transforms the bulk of cells into tumorigenic CD44(+)/CD24(low) cells with CSC properties, termed here as induced CSC-like-10A or iCSCL-10A cells. These reprogrammed cells display a malignant phenotype in culture and form tumors of multiple lineages when injected into immunocompromised mice. Compared with other transformed cell lines, cultured iCSCL-10A cells exhibit increased resistance to the chemotherapeutic compounds, Taxol and Actinomycin D, but higher susceptibility to the CSC-selective agent Salinomycin and the Pin1 inhibitor Juglone. Restored expression of the cyclin-dependent kinase inhibitor p16INK4a abrogated the CSC properties of iCSCL-10A cells, by inducing cellular senescence. This study provides some insight into the potential oncogenicity that may arise via cellular reprogramming, and could represent a valuable in vitro model for studying the phenotypic traits of CSCs per se.

Developmental changes in desensitisation of c-Fos expression induced by repeated maternal separation in pre-weaned mice.

Early-life stress has long-lasting effects on neuroendocrine and behaviour in adulthood. Maternal separation (MS) is used as a model of early-life stress and daily repeated MS (RMS) for 3 h during the first two postnatal weeks is widely used in rodent studies. However, it is not fully understood whether early-life animals desensitise/habituate to repeated stress. In the present study, we investigated the effects of daily RMS for 3 h and acute/single time MS (SMS) for 3 h on the plasma corticosterone level and c-Fos expression in the brain in mice at different postnatal ages. Mice were subjected to: (i) RMS from postnatal day (PND) 1 to 14 (RMS14); (ii) RMS from PND14 to 21 (RMS21); (iii) SMS on PND14 (SMS14); and (iv) SMS on PND21 (SMS21). Plasma corticosterone and c-Fos expression were examined on the final day in each experiment. The basal corticosterone levels in RMS14 and RMS21 were equal to those in respective age-matched controls. After the final separation, the levels were significantly increased and were comparable with those after SMS14 and SMS21, respectively. Histological analysis indicated that c-Fos expression significantly increased in many brain regions, including the paraventricular nucleus, prefrontal cortex, hippocampus, and basolateral and medial amygdale in both SMS14 and SMS21 mice. However, c-Fos expression in RMS14 mice significantly increased in many regions, whereas such increases were hardly seen in RMS21 mice. These results indicate that repeated early-life stress neither increases basal corticosterone, nor decreases the magnitude of the corticosterone response during the first three postnatal weeks, although desensitisation of c-Fos expression induced by repeated stress is changed during postnatal development.

Suppression of the let-7b microRNA pathway by DNA hypermethylation in infant acute lymphoblastic leukemia with MLL gene rearrangements.

MicroRNAs (miRNAs) regulate cell proliferation and differentiation by controlling the expression of proteins involved in many signaling pathways. Recent studies have shown that dysregulation of miRNA expression is associated with increased tumorigenicity and a poor prognosis in several types of cancers. The miRNA let-7b is one of the severely downregulated miRNAs in mixed-lineage leukemia (MLL)-rearranged acute lymphoblastic leukemia (ALL) patients. In vitro transfection of leukemogenic MLL fusion genes into human embryonic kidney-293 cells suppressed let-7b expression. In leukemic cells with an MLL fusion gene, the regulatory region for let-7b expression was hypermethylated, and its expression was partially recovered after culturing the cells with the demethylating agent 5-azacitidine. These results suggest that loss of let-7b expression may be one of the consequences of oncogenic MLL fusion proteins, and contributes to leukemogenesis possibly through the upregulation of let-7b-regulated target genes with leukemogenic potential in hematopoietic cells. The enforced expression of let-7b in ALL cell lines with an MLL fusion gene inhibited their growth, indicating the possible use of let-7b as a new therapeutic tool for refractory infant ALL with an MLL fusion gene.